Busca avançada
Ano de início
Entree
(Referência obtida automaticamente do Web of Science, por meio da informação sobre o financiamento pela FAPESP e o número do processo correspondente, incluída na publicação pelos autores.)

A calmodulin-like protein (LCALA) is a new Leishmania amazonensis candidate for telomere end-binding protein

Texto completo
Autor(es):
Mostrar menos -
Morea, Edna G. O. [1] ; Viviescas, Maria Alejandra [1] ; Fernandes, Carlos A. H. [2] ; Matioli, Fabio F. [2] ; Lira, Cristina B. B. [1] ; Fernandez, Maribel F. [3] ; Moraes, Barbara S. [4] ; da Silva, Marcelo S. [5] ; Storti, Camila B. [1] ; Fontes, Marcos R. M. [2] ; Cano, Maria Isabel N. [1]
Número total de Autores: 11
Afiliação do(s) autor(es):
[1] Sao Paulo State Univ UNESP, Biosci Inst, Genet Dept, BR-18618689 Botucatu, SP - Brazil
[2] Sao Paulo State Univ (UNESP), Biosci Inst, Biophys & Phys Dept, Botucatu, SP - Brazil
[3] ITPAC Porto Nacl SA, Inst Tocantinense Presidente Antonio Carlos LTDA, Porto Nacional, TO - Brazil
[4] PROAHSA Programa Estudos Avancados Adm Hosp & Sis, Sao Paulo - Brazil
[5] Butantan Inst, Ctr Toxins Immune Response & Cell Signaling CeTIC, Lab Especial Ciclo Celular, Sao Paulo, SP - Brazil
Número total de Afiliações: 5
Tipo de documento: Artigo Científico
Fonte: BIOCHIMICA ET BIOPHYSICA ACTA-GENERAL SUBJECTS; v. 1861, n. 11, A, p. 2583-2597, NOV 2017.
Citações Web of Science: 1
Resumo

Background: Leishmania spp. telomeres are composed of 5'-TTAGGG-3' repeats associated with proteins. We have previously identified LaRbp38 and LaRPA-1 as proteins that bind the G-rich telomeric strand. At that time, we had also partially characterized a protein: DNA complex, named LaGT1, but we could not identify its protein component. Methods and results: Using protein-DNA interaction and competition assays, we confirmed that LaGT1 is highly specific to the G-rich telomeric single-stranded DNA. Three protein bands, with LaGT1 activity, were isolated from affinity-purified protein extracts in-gel digested, and sequenced de novo using mass spectrometry analysis. In silico analysis of the digested peptide identified them as a putative calmodulin with sequences identical to the T. cruzi calmodulin. In the Leishmania genome, the calmodulin ortholog is present in three identical copies. We cloned and sequenced one of the gene copies, named it LCalA, and obtained the recombinant protein. Multiple sequence alignment and molecular modeling showed that LCalA shares homology to most eukaryotes calmodulin. In addition, we demonstrated that LCalA is nuclear, partially co-localizes with telomeres and binds in vivo the G-rich telomeric strand. Recombinant LCalA can bind specifically and with relative affinity to the G-rich telomeric single-strand and to a 3'G-overhang, and DNA binding is calcium dependent. Conclusions: We have described a novel candidate component of Leishmania telomeres, LCalA, a nuclear calmodulin that binds the G-rich telomeric strand with high specificity and relative affinity, in a calcium-dependent manner. (AU)

Processo FAPESP: 15/18641-8 - Caracterização molecular de RNAs teloméricos não codificadores em Leishmania major
Beneficiário:Maria Isabel Nogueira Cano
Linha de fomento: Auxílio à Pesquisa - Regular
Processo FAPESP: 12/50161-8 - Estudo da estrutura e função da chaperona Hsp90 com ênfase no seu papel em homeostase celular
Beneficiário:Carlos Henrique Inacio Ramos
Linha de fomento: Auxílio à Pesquisa - Temático